Profiling Urine Glycosylation of PSA and other Glyco-Biomarkers in Prostate Cance

描述(由申请人提供)：尿液是一种容易获得但在很大程度上被丢弃的泌尿生殖道癌生物标志物的潜在来源。癌症的生物标记物出现在尿液中，甚至来自肺癌和乳腺癌等远程癌症。我们观察到健康人尿中前列腺特异性抗原(PSA)和前列腺酸性磷酸酶(PAP)的显著水平，这与早期的一些发现一致。虽然PSA是目前前列腺癌(PC)的生物标志物，但其可靠性受到个体之间基线水平的差异以及各种可用的试剂盒测量结果不一致的影响。目前迫切需要建立一种具有临床应用价值的PSA(和PAP)检测方法，用于前列腺癌的早期发现、预后监测和正常细胞释放的PSA(和PAP)。PSA和PAP都是糖基化的，糖基化的变化与癌症密切相关。最近的报道表明，PSA的糖基化在癌症中发生了变化，但在不同的体液(PC的血清和健康的精液)中进行了测量，并且只在一个患者中有效。我们已经开发了一种方法(GlycoMatic)来分析浓缩蛋白质中个别位置的糖基化情况。我们将改进分离方法，以丰富PSA、PAP和其他可能在临床样本中检测到的前列腺特异性糖蛋白。我们将对照添加到健康人尿样中的标准和标准来测试该技术，获得可重复性测定的能力，以及糖蛋白在不太理想的条件下储存时的稳定性。为了消除可归因于遗传或表观遗传构成的任何变化，将使用60名正常志愿者的尿液建立个体PSA和PAP糖基化图谱的变异。关于年龄和族裔等变量的信息将被纳入分析，这些分析将产生于人口中有相当大的少数群体。将测定60例前列腺癌和良性前列腺增生症(BPH)患者的PSA和PAP糖基化图谱，并与健康志愿者的结果进行比较。获得用于临床试验的糖基化图谱的能力是一个新领域。因此，将建立计算方法，以显示每个疾病分类中个人内部数据的可靠性，并与疾病状况、Gleason评分和PSA血清水平相关。这项研究开辟了新的领域，并将为使用生物标记物蛋白的糖基化图谱在癌症的早期检测、预测和监测中的临床适用性奠定基础。它还将识别和区分任何因种族而产生的变化。前列腺特异性抗原(PSA)是用于前列腺癌(PC)早期发现和监测的一种成熟的检测方法。尽管如此，由于个体之间基础水平的差异，以及各种可用试剂盒的测量结果不一致，它的测定受到了阻碍。在癌症中产生的PSA被修饰(糖基化)，这项有限的临床试验将确定这些差异，并确定这些差异是否足够具体，以明确区分PC产生的PSA和正常情况下由健康前列腺产生的PSA。前列腺酸性磷酸酶，前列腺癌的早期糖蛋白生物标记物，也将进行类似的研究。分析将在尿液中进行，从而有可能对PC进行自我管理测试。

DESCRIPTION (provided by applicant): Urine represents an easily available yet largely discarded potential source of biomarkers of urogenital cancers. Biomarkers of cancer appear in urine, even from remote cancers such as lung and breast. We observed significant levels of both prostate specific antigen (PSA) and prostatic acid phosphatase (PAP) in the urine from healthy individuals, consistent with some earlier findings. While PSA is the current biomarker for prostate cancer (PC), its reliability is hampered by variations in the baseline level between individuals, and the inconsistency in measurements made by the variety of kits available. There is an urgent need for establishing a clinically pertinent assay for the characterization of PSA (and PAP) produced in PC, benign prostatic hyperplasia (BPH) and that released by healthy cells, one that can be reliably used for the early detection, prognosis and monitoring of prostate cancer. Both PSA and PAP are glycosylated and changes in glycosylation go hand-in-hand with cancer. Recent reports have shown that changes occur in the glycosylation of PSA in cancer but were measured in different body fluids (serum for PC and seminal fluid for healthy), and effectively in only one patient. We have developed a method (GlycoMatic) for profiling glycosylation at individual sites in enriched proteins. We will refine isolation approaches to enrich the PSA, PAP and such other prostate-specific glycoproteins as may be detected in clinical samples. We will test the technique against standards and standards spiked into urine samples from healthy individuals, the ability to get reproducible determinations, and the stability of the glycoproteins when stored under less than ideal conditions. To eliminate any changes attributable to genetic or epigenetic make-up, variations in the individual PSA and PAP glycosylation profiles will be established using urines from 60 normal volunteers. Information on variables such as age and ethnicity will be factored into the analyses which will result from a population with a significant minority component. The PSA and PAP glycosylation profiles will be determined for a cohort of 60 (of each) PC and benign prostatic hyperplasia (BPH) patients and compared to that from the healthy volunteers. The ability to obtain glycosylation profiles for clinical trials is a new field. Thus, computational methods will be established that show the reliability of the data within individuals, within each classification of the disease, and correlated with disease status, Gleason score and PSA serum levels. This research breaks new ground and will establish the clinical applicability for the use of glycosylation profiles of biomarker proteins in the early detection, prediction and monitoring of cancer. It will also identify and distinguish any changes due to ethnicity. Prostate specific antigen (PSA) is a well-established test for the early detection and monitoring of prostate cancer (PC). Despite this, its determination is hampered by variation in the basal level between individuals, and the inconsistency in measurements made by the variety of kits available. PSA produced in cancer is modified (glycosylated) and this limited clinical trial will establish the differences, and determine if these are specific enough to clearly distinguish PC produced PSA from that normally produced by a healthy prostate. Prostatic acid phosphatase, an earlier glycoprotein biomarker of prostate cancer, will be similarly studied. Analyses will be performed in urine, leading to the possibility of a self-administered test for PC.